Purification of DNA-dependent RNA polymerase Ii from Saccharomyces cerevisiae
Analytical Biochemistry 128(2): 294-301
ISSN/ISBN: 0003-2697 PMID: 6342468 DOI: 10.1016/0003-2697(83)90378-0
A rapid procedure for the purification of RNA polymerase II from Saccharomyces cerevisiae is described. Total RNA polymerase activity was solubilized from whole cells by sonication in 0.32 M (NH4)2SO4 and RNA polymerase II purified by polyethylenimine fractionation, ammonium sulfate precipitation, and chromatography on DEAE-cellulose, DEAE-Sephadex, and phosphocellulose. The procedure may be completed in 2.5 days and the resultant enzyme is judged to be greater than 90% pure.